Tuft cell heterogeneity in function, lineage, and structure in ileal inflammatory disease

项目摘要/摘要 炎症性肠病(IBD)及其亚型克罗恩病(CD)是由于缺乏 遗传易感个体对环境抗原的耐受性。纵向分析 已经确定了传染性寄生虫病和IBD之间的负相关， 这一现象被称为“卫生假说”。簇状细胞最近被鉴定为一种 感知和响应寄生虫感染的关键组件。我们的实验室最近 发现簇状细胞在微生物组中可能在谱系和功能上是不同的 反应和特定人群可能是减轻回肠炎性炎症的关键 疾病。在这里，我们提出了创新的实验和计算方法来剖析 簇状细胞谱系、结构和功能的异质性与免疫调节的关系 CD体内模型的反应。在目标1中，我们将进行单细胞测序，高 分辨率显微镜和功能分泌分析以了解谱系结构功能 在小鼠和肠样体模型中不同簇状细胞群之间的关系。在目标2中，我们利用 多重成像、细胞计数、飞行时间(CyTOF)和细胞因子分析以探测 利用几种化学和遗传模型的下游簇状2型反应网络 径路消融。通过我们的发现，我们的目标是为当前的 了解绒毛细胞生物学和肠道上皮-微生物组串扰。最终， 这项研究将使我们能够诊断、分层，并最终治疗IBD患者并扩大 这种复杂的、多因素疾病的靶点。

PROJECT SUMMARY/ABSTRACT Inflammatory Bowel Disease (IBD) and its subtype Crohn’s disease (CD) arise due to a loss of tolerance to environmental antigens in genetically susceptible individuals. Longitudinal analysis has identified an inverse correlation between rates of communicable parasitic diseases and IBD, a phenomenon termed the “hygiene hypothesis.” Tuft cells have recently been identified as a critical component for sensing and responding to parasitic infections. Our lab has recently identified that tuft cells may be heterogeneous in their lineage and function in microbiome response, and specific populations may be critical for mitigating inflammation in ileal inflammatory disease. Here, we propose innovative experimental and computational approaches to dissect the heterogeneity of tuft cell lineage, structure, and function as they relate to regulating immune responses in in vivo models of CD. In aim 1, we will conduct single-cell sequencing, high resolution microscopy, and functional secretory assays to understand lineage-structure-function relationships in different tuft cell populations in mouse and enteroid models. In aim 2, we utilize multiplex imaging, Cytometry Time-of-Flight (CyTOF), and cytokine profiling to probe the downstream tuft-type 2 response network by utilizing several chemical and genetic models of pathway ablation. Through our findings, we aim to make significant contributions to the current understanding of tuft cell biology and epithelial-microbiome crosstalk in the intestine. Ultimately, this research will allow us to diagnose, stratify, and, ultimately, treat patients with IBD and expand the pool of targets in this complex, multifactorial disease.